Development of a panel of seven duplex real-time PCR assays for detecting 13 streptococcal superantigens

Annals of Clinical Microbiology and Antimicrobials

Research

Shujuan Cui¹  Quanyi Wang¹  Huijie Liang¹  Peng Yang¹  Daitao Zhang¹  Xiaomin Peng¹  Xuping Zhu²  Junbin Shao² 

[1] Institute for Infectious Disease and Endemic Disease Control, Beijing Center for Disease Prevention and Control (CDC), Beijing Research Center for Preventive Medicine; Capital Medical University School of Public Health, No.16 He Pingli Middle Street, 100013, Dongcheng District, Beijing, China;Shanghai ZJ Biotech Co. Ltd., No. 188 Xinjunhuan Road, Pujiang High-tech Park Caohejing Development area, 201114, Shanghai, China;

关键词: Streptococcal;    Superantigens;    Duplex real-time PCR;   

DOI  :  10.1186/1476-0711-12-18

 received in 2013-03-19, accepted in 2013-07-27,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundStreptococcal superantigens (SAgs) are the major virulence factors of infection in humans for group A Streptococcus (GAS) bacteria. A panel consisting of seven duplex real-time PCR assays was developed to simultaneously detect 13 streptococcal SAgs and one internal control which may be important in the control of GAS-mediated diseases.MethodsPrimer and probe sequences were selected based on the highly conserved region from an alignment of nucleotide sequences of the 13 streptococcal SAgs. The reaction conditions of the duplex real-time PCR were optimized and the specificity of the duplex assays was evaluated using SAg positive strains. The limit of detection of the duplex assays was determined by using 10-fold serial dilutions of the DNA of 13 streptococcal SAgs and compared to a conventional polymerase chain reaction (PCR) method for evaluating the duplex assays sensitivity.ResultsUsing the duplex assays, we were able to differentiate between 13 SAgs from Streptococcus strains and other non-Streptococcus bacteria without cross-reaction. On the other hand, the limit of detection of the duplex assays was at least one or two log dilutions lower than that of the conventional PCR.ConclusionsThe panel was highly specific (100%) and the limit of detection of these duplex groups was at least ten times lower than that obtained by using a conventional PCR method.

【 授权许可】

Unknown   
© Yang et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.